Rho/Rho激酶信号通路与糖尿病肾病

糖尿病肾病(DN)是糖尿病重要的微血管并发症之一,其发病机制是多种因素共同作用的结果.近年来发现Rho/Rho激酶(Rho/ROCK)信号通路与多种组织重构如血管、心肌、肾纤维化等相关.高糖能够刺激肾脏细胞内Rho/ROCK信号通路的激活,后者通过调控转录因子的DNA结合活性上调多种基因的表达,从而引起各种炎性反应及肾小球纤维化,而应用此信号通路选择性抑制剂后可明显改善DN的发生和发展,从而认为Rho/ROCK信号转导通路可能在DN的发病机制中起关键作用.     

Rho/ROCK信号通路在慢性充血性心力衰竭中的作用

Rho/Rho相关的卷曲蛋白激酶（ROCK）信号通路的关键信号分子包括Rho、ROCK及其主要效应分子肌球蛋白轻链磷酸酶（MLCP）、肌球蛋白轻链激酶（MLCK）、肌球蛋白轻链（MLC）等,该信号通路激活后可产生多种生物效应,如肌动蛋白细胞骨架形成、细胞收缩、钙敏化等,且这些效应与心力衰竭的发展过程有关.目前,对于Rho/ROCK信号通路的研究主要集中在心、脑血管系统中,如缺血性卒中、高血压、冠心病等.越来越多的研究表明,Rho/ROCK信号通路的关键分子在慢性充血性心力衰竭（CHF）中存在异常表达与活化,提示该信号转导通路在CHF发病机制中具有重要的作用.本文结合文献就Rho/ROCK信号通路在CHF过程中对细胞舒缩功能、交感兴奋、细胞凋亡等的作用综述如下.     

Rho／Rho激酶在肺癌中的作用及其潜在治疗靶点

Rho蛋白在细胞内起到分子开关的作用,Rho／ROCK信号通路在细胞的粘附、变形、迁移、增殖、凋亡等多种行为方面起到重要调控作用。Rho蛋白的异常表达或激活被发现与肺癌细胞生物学特征和患者预后等有显著相关性,并可能成为潜在治疗靶点。现就Rho／ROCK信号通路在肺癌中的作用及其潜在治疗作用做一综述。     

Rho/Rho激酶信号通路与缺血性脑血管病

Rho/Rho激酶信号通路是体内普遍存在的一条信号通路,它通过调节细胞内肌动蛋白骨架的聚合状态而扮演着"分子开关"角色,参与多种细胞功能.Rho/Rho激酶信号通路在缺血性脑血管病的危险因素、发病机制和病理生理学过程中发挥着重要作用,抑制Rho/Rho激酶信号通路能够取得显著的神经保护作用.     

Rho/Rho激酶信号通路与冠心病

Rho/Rho激酶信号通路是体内普通存在的一条信号转导通路,它通过调节细胞内肌动蛋白骨架的聚合状态参与多种细胞功能,包括细胞收缩、迁移、黏附、增殖、凋亡及基因表达等。Rho/Rho激酶信号通路的异常激活在冠心病的发病机制和病理生理中发挥了重要作用,对此信号转导通路的研究可以为冠心病的预防和治疗提供新的靶点。现就Rho/Rho激酶信号通路的特征及其与冠心病的关系作一综述。     

Rho/ROCK与神经系统疾病

作为一种分子开关,Rho可在无活性的GDP结合形式和有活性的GTP结合形式之间转换,传递信号给下游底物Rho激酶(ROCK),介导特殊的生物学效应,参与多种神经系统疾病,如缺血性卒中、脑血管痉挛、脊髓损伤后轴突再生的病理学过程.应用ROCK抑制剂治疗神经系统疾病已取得满意效果.Rho/ROCK有望成为一个新的治疗靶点.     

Rho激酶在心血管疾病中新进展及Rho激酶抑制剂的应用

Rho激酶(ROCK)被认为是治疗心血管疾病的新靶点。本文主要简述RhoA/ROCK通路在调节脉管系统的功能,并探讨其在心血管疾病发病中的作用机制。     

Rho激酶与心血管疾病

<正>1996年,人们发现Rho激酶(Rho associated kinase,ROCK)是Rho的下游效应器之一,随着研究的深入,发现其参与了血管平滑肌细胞、内皮细胞、外膜细胞、心肌细胞的形态、收缩、迁移、增殖、凋亡、基因转录等多种病理生理改变,它的异常活化与心血管疾病的发病机制密切相关。近年来关于ROCK的研究主要集中在心血管系统方面,主要因为:(1)Rho/ROCK通路参与的各种细胞功能在心血管疾病的发     

Rho相关蛋白激酶与动脉粥样硬化关系的研究进展

<正>动脉粥样硬化(AS)是一种累及全身大血管壁的慢性炎性反应和脂质沉积过程,一些危险因素(如高血糖、血脂异常以及高血压等)可以激活多种信号通路参与AS的发展,Rho A/Rho相关蛋白激酶(ROCK)通路就是其中之一。越来越多的证据表明,RhoA/ROCK通路参与AS过程的许多步骤,ROCK可能成为AS的潜在治疗靶点,了解Rho/ROCK通路在AS进展过程中的作用对AS的防治有重要意义^[1]。     

Rho/Rho激酶信号通路与低氧所致肺纤维化

小G蛋白Rho参与了细胞黏附、迁移、生长、细胞收缩及细胞分裂等多种生物学行为.近来研究显示Rho/Rho激酶信号通路的异常活化参与了肺部疾病的发生,如肺动脉高压和肺纤维化.低氧作为一种潜在的促纤维化因素主要通过促内皮细胞凋亡、血管生成及炎症反应的调节来参与纤维化的发生.且Rho/Rho激酶信号通路与低氧所致肺纤维化的过程中所涉及主要细胞因子都有着直接或IhJ接的关系.因此,本文就Rho/Rho激酶信号通路的生物学特征及其与低氧致肺纤维化过程中细胞凶子的关系作一综述.     

RhoA在肝纤维化中的作用机制

肝纤维化发生的关键在于肝星状细胞（HSC）转化为成纤维母细胞及活化后细胞外基质（ECM）的异常表达。回顾了Rho/ROCK信号通路在HSC的激活中的重要作用。总结了RhoA,作为Rho/ROCK信号通路上小G蛋白家庭成员之一,且作为一种重要的调节肌动蛋白细胞骨架和细胞形态异质性的细胞因子,参与肝纤维化的进程。指出RhoA在肝脏纤维化发生发展过程中起着重要的调控作用,归纳了RhoA通过调节肌动蛋白应力纤维的装配,并通过级联放大效应影响细胞基质的合成和收缩,最终影响肝纤维化的进展。就RhoA在肝纤维化过程中的机制以及RhoA作为潜在的治疗靶点作一综述。     

microRNA-124与糖尿病肾脏疾病

microRNA(miRNA)广泛存在于真核生物体内,参与蛋白质的转录和转录后的调控.miRNA具有高度进化保守性及组织特异性,在生物的生长发育、细胞分化、疾病的发展过程中起作用.miRNA-124是microRNAs家族的重要成员,近年研究显示,miRNA-124能够作用于整合素,进而影响肾脏纤维化进程,可能作用于Rho/Rho相关蛋白激酶信号通路,起到保护肾小球滤过屏障的作用,能够作用于信号转导与转录激活因子3、Toll样受体、磷脂酰肌醇3激酶/蛋白激酶B信号通路调节炎性反应,可能成为糖尿病肾脏疾病诊断和治疗的新靶点.     

Long-term consumption of Western diet contributes to endothelial dysfunction and aortic remodeling in rats: Implication of Rho-kinase signaling

Western diet (WD), rich in saturated fat and sugars, has become a risk factor for obesity and metabolic syndrome, however, its effect on endothelial function and vascular remodeling is not fully elucidated. Recent evidence suggests cross-talk between Rho kinase (ROCK) pathway and cardiovascular system. We aimed to investigate the effect of WD on aortic remodeling and the contribution of ROCK signaling. Eight week old male Sprague-Dawley rats were fed either standard chow diet (SD) or high fructose/ high-fat diet, typically as in WD. After 42 weeks, WD-fed rats showed hyperglycemia, dyslipidemia, and hypertension without marked weight gain, compared to SD-fed rats. Significant up-regulation of ROCK-1 and ?2, along with a decline in eNOS expression were found in the aortic tissue of WD-fed rats. Additionally, WD-fed rats displayed oxidative stress and fibrosis in their aortic tissues versus controls. Our findings suggest that long-term feeding of WD contributes to endothelial dysfunction and aortic remodeling in adult male rats. ROCK activation seems to be involved in WD-related vascular disorders and may represent a promising therapeutic target.     

非对称性二甲基精氨酸通过Rho／ROCK信号通路介导大鼠血管平滑肌细胞迁移

目的探讨非对称性二甲基精氨酸（Asymmetric Dimethylarginine,ADMA）对血管平滑肌细胞（vascular smooth muscle cells,VSMCs）迁移和形态变化的影响,并探索Rho／ROCK和MAPK信号转导通路在其中的作用。方法原代培养大鼠VSMCs,并通过转染二甲基精氨酸二甲胺水解酶hDDAH2过表达载体,L-精氨酸（1mM）或ROCK抑制剂Y27632等预处理,观察细胞RhoA蛋白与底物rhotekin蛋白结合程度,ROCK底物MYPT-1磷酸化程度、VSMC迁移能力和细胞骨架及黏着斑定位情况。结果（1）ADMA诱导平滑肌细胞Rho/OCK信号转导通路;（2）Rho/OCK和ERK信号通路交联介导ADMA对平滑肌细胞迁移能力的诱导作用;（3）ADMA通过Rho/OCK诱导平滑肌细胞骨架无序排列、黏着斑定位改变;（4）L-精氨酸对上述变化有逆转作用。结论ADMA通过Rho/OCK和ERKl／2信号交联诱导VSMC迁移和表型转化,而L-精氨酸逆转ADMA引起的改变。     

高糖通过Rho/ROCK信号通路诱导人肾小球系膜细胞的炎症反应及纤维化

目的 探讨Rho/ROCK信号通路在高糖诱导的人肾小球系膜细胞(HMCs)炎症反应及纤维化中的作用.方法 将传代培养的HMCs同步化后分组:(1)正常糖浓度对照组(NG,5.5 mmol/L葡萄糖);(2)高糖组(HG,30 mmol/L葡萄糖);(3)甘露醇渗透压对照组(Man,5.5 mmol/L葡萄糖+24.5 mmol/L甘露醇);(4)NG+Y-27632(10 μmmol/L)组;(5)HG+Y-27632(10 μmmol/L)组,培养12、24、36、48、72 h后收集上清及细胞,用Western印迹检测RhoA蛋白的活化,用实时PCR检测细胞中RhoA、ROCK-Ⅰ、结缔组织生长因子(CTGF)、肿瘤坏死因子α(TNF-α)mRNA浓度的变化,用ELISA方法检测上清中纤维连接蛋白(FN)、CTGF、TNF-α的蛋白含量.结果 (1)高糖刺激HMCs的RhoA活化,于30 min即可出现活性升高,1 h达到高峰,之后活化的RhoA表达逐渐下降(P=0.02).(2)高糖培养下的HMCsRhoA、ROCK-Ⅰ、CTGF、TNF-α mRNA的表达较NG组明显升高(P＜0.05),并有一定的时间依赖性,Man组与NG组相比差异无统计学意义(P＞0.05).(3)经Y-27632预处理后,在正常糖和高糖浓度培养24 h或48 h后,NG+Y-27632组和HG+Y-27632组与未处理组相比RhoA、ROCK-Ⅰ、CTGF、TNF-α mRNA的表达明显下降(P＜0.01).(4)高糖呈时间依赖方式增加HMCs的FN、CTGF、TNF-α分泌(P＜0.05).(5)经Y-27632预处理,继续培养12、24、36、48、72 h后NG组和HG组中FN、CTGF、TNF-α蛋白的分泌较处理前明显降低(P＜0.05).结论 高糖可通过Rho/ROCK信号通路介导HMCs的炎症反应和纤维化,抑制此通路可作为减缓糖尿病肾病发生发展的潜在靶点.

Abstract：

Objective To investigate the role of Rho/ROCK signaling pathway in the process of human mesangial cells (HMCs) inflammation and fibrosis induced by high glucose. Methods Synchronized HMCs were divided into following groups: ( 1 ) Normal glucose control group ( NG, 5.5 mmol/L glucose); ( 2 ) High glucose group ( HG, 30 mmol/L glucose); (3) Mannitol group( Man,5.5 mmol/L glucose+ 24.5 mmol/L mannitol); (4) NG +Y-27632 group( 10 μ mmol/L Y-27632 ); ( 5 ) HG Y-27632 group ( 10 μmmol/L Y-27632 ). The supernatant and cells were collected at 0,12,24,36,48, and 72 h. Western blot was used to detect the active RhoA and total RhoA,while RhoA, ROCK-Ⅰ, CTGF, and TNF-α mRNA expressions were determined with realtime PGR method in the cells, then ELISA method was used to check protein levels of FN, CTGF, and TNF-α in the supernatant. Results ( 1 ) RhoA activation was stimulated after treatment for with 30 mmol/L glucose, peaked at 1 h, and then decreased ( P = 0. 02). (2) RhoA, ROCK-Ⅰ, CTGF, and TNF-α mRNA expressions in HMC cultured under high glucose were higher than those in the normal group ( P ＜ 0.05 ), and there was certain time-dependence. Besides, there was no statistical significance between Man and NG groups( P＞0. 05 ). ( 3 ) After Y-27632 pretreatment and being cultured with normal glucose and high glucose for24 h or48 h, RhoA, ROCK-Ⅰ, CTGF, and TNF-α mRNA expressions were significantly decreased ( P＜0.01 ) as compared with groups without treatment. (4) High glucose increased FN, CTGF,and TNF-α protein secretion of HMC in a time-dependent manner( P＜0. 05 ). ( 5 ) After Y-27632 pretreatment and being cultured with normal and high glucose for 12,24,36,48,72 h, FN, CTGF, and TNF-α protein secretions were significantly reduced( P＜0.05 ). Conclusion Rho/ROCK signaling pathway may mediate inflammation and fibrosis induced by high glucose in HMCs, supporting a potential role for inhibitors of Rho/ROCK in the treatment of diabetic nephropathy.     

Rho/Rho kinase is a key enzyme system involved in the angiotensin II signaling pathway of liver fibrosis and steatosis

BACKGROUND AND AIM: The molecular mechanisms underlying the involvement of the renin-angiotensin system in hepatic fibrosis are unclear. Recently, it was reported that a Rho kinase inhibitor prevented fibrosis of various tissues and that the Rho/Rho kinase pathway was involved in the renin-angiotensin system of vascular smooth muscle cells. In this study, the involvement of the Rho/Rho kinase pathway on angiotensin II signaling in liver fibrogenesis and generation of steatosis was investigated. METHODS: Rats were fed a choline-deficient/L-amino acid-defined (CDAA) diet continuously and treated with a Rho kinase inhibitor, Y-27632, and an angiotensin II receptor blocker, TCV-116. Liver histology and hepatic stellate cell activation were analyzed. Free radical production was detected by 4-hydroxynonenal and 8-hydroxy-2'-deoxyguanosine immunostaining and the expression of tumor necrosis factor-alpha was examined. Isolated hepatic stellate cells were pretreated with a Rho kinase inhibitor, Y-27632, or an angiotensin II receptor blocker, CV-11974, and stimulated with angiotensin II, and mRNA expression of transforming growth factor-beta and alpha-smooth muscle actin was analyzed. RESULTS: Both the angiotensin II receptor blocker and the Rho kinase inhibitor improved fibrosis and steatosis of the liver in CDAA-fed rats. The increase in the number of hepatocytes positive for 4-hydroxynonenal and 8-hydroxy-2'-deoxyguanosine in CDAA-fed rats was significantly prevented by the angiotensin II receptor blocker and the Rho kinase inhibitor. The levels of tumor necrosis factor-alpha mRNA in the liver of CDAA-fed rats were significantly increased and this increase was significantly inhibited by treatment with the angiotensin II receptor blocker and the Rho kinase inhibitor. mRNA expression of transforming growth factor-beta and alpha-smooth muscle actin stimulated by angiotensin II was also significantly suppressed by these two drugs. CONCLUSION: These results suggest that the Rho/Rho kinase pathway is at least partly involved in the renin-angiotensin system and plays an important role in hepatic fibrosis and steatosis.     

小G蛋白Rho与肾间质纤维化

肾间质纤维化是所有慢性进行性肾脏疾病进展到终末期肾衰竭的共同形态学特点。Rho蛋白作为重要的细胞内信号分子,调节细胞的多种行为和功能,包括细胞生长,细胞骨架的重组,细胞移行和增殖等。近年研究发现,Rho蛋白及相关信号通路在肾间质纤维化过程中起着重要作用。通过作用于该通路从而达到治疗或逆转肾功能进行性损害的目的,可成为一个新的研究方向。     

Long-term diabetic complications may be ameliorated by targeting Rho kinase

This review addresses the roles of Rho/Rho-kinase (ROCK) pathway in the pathogenesis of diabetes complications. Diabetes can cause many serious complications and can result in physical disability or even increased mortality. However, there are not many effective ways to treat these complications. The small guanosine-5'-triphosphate-binding protein Rho and its downstream target Rho-kinase mediate important cellular functions, such as cell morphology, motility, secretion, proliferation, and gene expression. Recently, the Rho/Rho-kinase pathway has attracted a great deal of attention in diabetes-related research. These studies have provided evidence that the activity and gene expression of Rho-kinase are upregulated in some tissues in animal models of type 1 or type 2 diabetes and in cell lines cultured with high concentrations of glucose. Inhibitors of Rho-kinase could prevent or ameliorate the pathological changes in diabetic complications. The inhibitory effects of statins on the Rho/Rho-kinase signalling pathway may also play a role in the prevention of diabetic complications. However, the precise molecular mechanism by which the Rho/Roh-kinase pathway participates in the development or progression of diabetic complications has not been extensively investigated. This article evaluates the relationship between Rho/Roh-kinase activation and diabetic complications, as well as the roles of Roh-kinase inhibitors and statins in the complications of diabetes, with the objective of providing a novel target for the treatment of long-term diabetic complications.